EP1453861A2 - Polynukleotid, protein und varianten, verwickelt in die synaptogenese, und deren diagnostische und therapeutische anwendungen - Google Patents

Polynukleotid, protein und varianten, verwickelt in die synaptogenese, und deren diagnostische und therapeutische anwendungen

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Publication number
EP1453861A2
EP1453861A2 EP02801090A EP02801090A EP1453861A2 EP 1453861 A2 EP1453861 A2 EP 1453861A2 EP 02801090 A EP02801090 A EP 02801090A EP 02801090 A EP02801090 A EP 02801090A EP 1453861 A2 EP1453861 A2 EP 1453861A2
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EP
European Patent Office
Prior art keywords
protein
seq
polypeptide
polynucleotide
mutation
Prior art date
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EP02801090A
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English (en)
French (fr)
Inventor
Thomas Bourgeron
Stéphane JAMAIN
Hélène QUACH
Catalina Betancur
Marion Leboyer
Christopher Gillberg
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Assistance Publique Hopitaux de Paris APHP
Institut National de la Sante et de la Recherche Medicale INSERM
Institut Pasteur
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Assistance Publique Hopitaux de Paris APHP
Institut National de la Sante et de la Recherche Medicale INSERM
Institut Pasteur
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Publication of EP1453861A2 publication Critical patent/EP1453861A2/de
Withdrawn legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system

Definitions

  • the present invention relates to the identification of a human gene coding for a protein involved in synaptogenesis and of its murine ortholog, and whose mutation is associated in humans with the development of neurological diseases and / or with a predisposition to the development of mental disorders or psychiatric illnesses such as autism and Asperger's Syndrome.
  • the invention also relates to diagnostic and therapeutic applications related to the identification of the gene and its mutations.
  • the invention also relates to diagnostic and therapeutic applications linked to the identification of the involvement of a defect in a protein of the neuroligin family in the development of mental disorders or psychiatric diseases such as autism and the Syndrome of Asperger. b) Brief description of the prior art
  • Autism is a disease which affects approximately one child in 1000 and mainly boys (from 4 to 23 boys for a girl depending on the clinical criteria chosen). The clinical symptoms of autism are described in the book DSM-IV-TR TM (Diagnostic and statistical manual of mental disorders, 2000, pages 70-75).
  • Neuroligins are cell adhesion proteins which alone can trigger synaptogenesis, i.e. the formation of synapses. (Scheiffele et a /., Cell, 2000, 101: 657-669). Neuroligins NL1, NL2 and NL3 were originally cloned in rats (Ichtchenko et al., Cell., 1995, 81 (3): 435-43; Ichtchenko et al., J. Biol. Chem., 1996, 271 ( 5): 2676-82). The neuroligins HNL1 and HNL2 are located on autosomes (3q26 and 17p13).
  • HNL4X Human Neuroligline-4
  • Bollinger et al. Biochem. J., 2001, 356: 581-588
  • the LOCUSLINK TM database http://www.ncbi.nim.nih.gov) dated September 13, 2001 provides under the accession numbers KIAA1260 and KIAA0951 incomplete sequences of a neuroligin gene the function of which is also unknown.
  • All neuroligins have an extracellular domain homologous to Acetylcholine esterase (ACHE).
  • ACHE Acetylcholine esterase
  • the neuroligins interact with the ⁇ -neurexins (Ichtchenko et al., Cell., 1995, 81 (3): 435-43). This interaction can be modulated by the ACHE itself (Grifman et al., Proc. Natl. Acad. Sci. USA, 1998, 95 (23): 13935-40).
  • ACHE Acetylcholine esterase
  • the present invention relates to the identification of human genes and their murine ortholog coding for a protein involved in synaptogenesis and whose mutation is associated in humans with the development of neurological diseases and / or with a predisposition to the development of mental disorders or psychiatric illnesses such as autism and Asperger's Syndrome.
  • the present invention relates to an isolated or purified polynucleotide coding for a polypeptide involved, in its wild form, in synaptogenesis.
  • the polynucleotide of the present invention is characterized in that at least one mutation in the nucleic acid sequence of said polynucleotide is associated with the development of neurological diseases and / or with a predisposition to the development of mental disorders or psychiatric diseases.
  • the present invention relates to a polynucleotide encoding a protein belonging to the family of human Neuroligins (HNLs) and more particularly the protein HNL4X (previously called HNL4) and its functional counterpart HNL4Y (previously called HNL5) coded by a chromosome gene.
  • HNLs human Neuroligins
  • HNL4X previously called HNL4X
  • HNL4Y previously called HNL5
  • the present invention also relates to a polynucleotide encoding the mouse protein MNL4 orthologue of the proteins HNL4X and HNL4Y.
  • the present invention relates to an isolated or purified polypeptide, characterized in that it is coded by a polynucleotide as defined above. More particularly, the polypeptide according to the present invention is characterized in that it is involved in synaptogenesis, and in that the presence of at least one mutation in the amino acid sequence of said polypeptide is associated with a predisposition to the development of mental disorders or psychiatric illnesses.
  • the present invention provides a method for detecting biochemical disorders altering the formation of synapses, and / or a predisposition to the development of psychiatric pathologies and / or a mental illness, comprising at least one of the following steps:
  • kits for the detection of biochemical disorders altering the formation of synapses, and / or a predisposition to the development of psychiatric pathologies and / or a mental illness, and / or for the diagnosis of a mental illness comprising at least one of the elements chosen from the group consisting of: a probe, an antibody, a reagent and a solid support allowing: i) the detection of a mutation in the sequence of a polynucleotide as defined above, in the sequence of a fragment of said polynucleotide or in the sequence of a messenger RNA of said polynucleotide; and / or ii) measuring the biological activity of a polypeptide as defined above or the interaction thereof with one of its protein partners.
  • the invention also relates to the use of a non-mutated polynucleotide encoding a protein involved in its wild form in synaptogenesis for the treatment or prevention of biochemical pathologies or mental diseases.
  • the invention also relates to the use of an unmutated polypeptide involved in its wild form in synaptogenesis for the treatment or prevention of biochemical pathologies or mental diseases.
  • the present invention also provides a method for sorting molecules making it possible to modulate the biological activity of the polypeptide encoded by the polynucleotide defined above or the biological activity of the polypeptide defined above, comprising: a) bringing said polypeptide or a recombinant cell containing it with a molecule capable of modulating its biological activity; b) measuring the biological activity of said polypeptide or its interaction with one of its protein partners; and c) evaluating the activity measured in step b) relative to a measurement of the biological activity of said polypeptide in the absence of said molecule.
  • Another object of the present invention is a method of treating a mental or neurological disease comprising the insertion into at least a portion of the cells of a patient of a patient of a polynucleotide encoding a polypeptide as defined above.
  • the present invention also provides a method of transforming stem cells of a patient with a mutation of a gene coding for a protein involved in synaptogenesis, characterized by a) the use of stem cells of said patient; b) the insertion into the genome of said stem cells of a polynucleotide as defined above; and c) reimplantation in the patient of cells transformed according to step b).
  • the invention further relates to a cloning or expression vector comprising one of the polynucleotides of the invention or a fragment thereof; a host cell containing a polynucleotide and / or a vector according to the invention; or purified monoclonal or polyclonal antibodies specifically recognizing at least one of the polynucleotides of the invention and / or at least one of the polypeptides of the invention.
  • the invention also relates to a composition containing at least one element chosen from the group consisting of a) a polypeptide, b) a polynucleotide, c) a vector, d) a host cell or e) an antibody, and a pharmaceutically acceptable vehicle.
  • Figure 1 shows the region of chromosome Xp22.3 containing the HNL4X gene.
  • Figure 2 shows a protein alignment of human Neuroligins.
  • Figure 3 is a diagram showing the protein architecture of a synapse with the location and known partners of Neuroligins.
  • Figures 4A and 4B represent a diagram which shows the chromosomal location and the evolution of the HNL4X and HNL4Y genes.
  • Figure 5 is a diagram showing the genomic structure of human Neuroligins genes.
  • Figures 6A and 6B show the result of the analysis by SSCP (Single Strand
  • Conformational Polymorphism of a mutation in the gene coding for the protein HNL4X as well as the sequence of this mutation.
  • Figure 7 is a diagram showing the location of the HNL4X protein and the effects of the mutation on HNL4X.
  • Figure 8A shows the genomic sequence (SEQ ID NO: 1) of the human gene
  • Figure 8B shows the nucleic acid sequence (SEQ ID NO: 16) of the exon
  • Figure 9 shows the complementary DNA sequence (SEQ ID NO: 2) of the human HNL4X wild type gene (not mutated).
  • Figure 10 shows the amino acid sequence (SEQ ID NO: 3) of the human protein HNL4X wild (not mutated).
  • Figure 11A shows the genomic sequence (SEQ ID NO: 4) of the human gene
  • Figure 11B shows the nucleic acid sequence (SEQ ID NO: 17) of exon 1 of Figure 11A.
  • Figure 12 shows the complementary DNA sequence (cDNA) (SEQ ID NO: 5) of the wild-type human HNL4 Y gene.
  • Figure 13 shows the amino acid sequence (SEQ ID NO: 6) of the human protein H NL4Y wild.
  • Figure 14 shows the complementary DNA sequence (cDNA) (SEQ ID NO: 7) of an alternative transcript of the wild-type human HNL4 Y gene.
  • Figure 15 represents the amino acid sequence (SEQ ID NO: 8) corresponding to the alternative sequence of Figure 14.
  • Figure 16 shows the amino acid sequence (SEQ ID NO: 9) of the mutated human protein HNL4X.
  • Figure 17 shows the chromosomal location of the HNL3, HNL4X and
  • HNL4Y and pedigree of a family with a mutation in HNL4X in an autistic boy and a boy with Asperger's syndrome are known in the art.
  • Figure 18 is a diagram showing the conservation of HNL3 mutations.
  • Figure 19A shows the nucleic acid sequence of MNL4 cDNA (SEQ ID NO: 1
  • Figure 19B shows the amino acid sequence of the MNL4 protein (SEQ ID NO: 1
  • Figure 20 is a diagram showing the genomic structure of the HNL1, HNL2 and HNL3 genes, as well as the location of the mutations observed in HNL3.
  • Figure 21 is a diagram showing the genomic structure of HNL4X and HNL4Y.
  • Figure 22 shows a portion of the amino acid sequence (SEQ ID NO: 10) of the HNL3 protein mutated at position 451.
  • Figure 23 shows another portion of the amino acid sequence (SEQ ID NO: 11) of the HNL3 protein mutated at position 796.
  • Figure 24 shows the complementary DNA sequence (cDNA) (SEQ ID NO: 12) of the wild-type HNL3 transcript.
  • Figure 25 shows the complementary DNA sequence (cDNA) (SEQ ID NO: 13) of the mutated HNL3 transcript.
  • Figure 26 shows the amino acid sequence (SEQ ID NO: 14) of the human protein HNL3 wild.
  • Figure 27 shows the amino acid sequence (SEQ ID NO: 15) of the mutated human protein HNL3.
  • the originality of the present invention relates to the identification of the genomic sequence of the HNL4X gene located in Xp22.3 and of a functional homolog HNL4Y placed on the Y chromosome located in Yq11.22 as well as their murine ortholog MLN4.
  • the invention also relates to the identification of the involvement of synaptogenesis proteins, in particular HNL3 and HNL4 in the development of mental disorders or psychiatric diseases such as autism.
  • the present invention relates to an isolated or purified polypeptide, which, in its wild form (ie non-mutated), is involved in synaptogenesis, of which at least one mutation in the amino acid sequence is associated with the development of neurological diseases and / or a predisposition to the development of mental disorders or psychiatric diseases.
  • “Mental disorders or psychiatric illnesses” means illnesses such as autism, Asperger's syndrome, schizophrenia and ADHD (Attention Deficit Hyperactivity Disorder) syndrome.
  • the polypeptide consists of a cell adhesion protein, more preferably a protein belonging to the family of human neuroligins and even more preferably, the polypeptide consists of the protein HNL3, HNL4X or the protein HNL4Y.
  • the polypeptide when it is HNL3, it comprises an amino acid sequence according to SEQ ID NO: 14 and the sequences of at least 20, at least 50 and at least 100 consecutive or more derived amino acids of SEQ ID NO: 14.
  • polypeptide of the invention comprises a sequence chosen from the group consisting of: SEQ ID NO: 3, SEQ ID NO: 6, SEQ ID NO: 8, and the sequences at least 20, at least 50 and at least 100 consecutive or more amino acids derived from SEQ ID NO: 3, SEQ ID NO: 6 or SEQ ID NO: 8.
  • the invention also relates to the "mutated" polypeptides and the polypeptides "derived” from the wild-type protein, preferably a neuroligin such as HNL3, HNL4X or HNL4Y.
  • polypeptide derived from a wild protein or “variant” of a wild protein is meant all peptides which have a peptide sequence substantially identical, at least in part, to the peptide sequence of the wild protein. They may, for example, be chemically modified polypeptides having a peptide sequence 100% identical to a portion of the wild-type protein. They may also be hybrid polypeptides having a first portion 100% identical to a first portion of the wild protein and a second portion in no way / partially identical to a second portion of the wild protein. They may also be polypeptides having total / partial homology with a portion of the wild-type protein.
  • mutant polypeptides derived from a wild protein all the peptides which have been obtained following a modification of said wild protein, whether it is a modification by addition, deletion or substitution of one or more of the amino acids of the wild protein. It can also be a modification brought about by the addition of carbon chains attached to at least one of the amino acids of the wild protein or to at least one of the amino acids of peptides for which there is a substitution or a modification of one of the amino acids compared to wild protein. More particularly, the present invention covers peptides which are derived from the human protein HNL3, HNL4X or HNL4Y. According to a preferred embodiment, and in the case where the polypeptide is a
  • HNL3 mutated according to the present invention, it has SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 15, and a sequence of at least 20, at least 50 and at least 100 consecutive or more amino acids derived from SEQ ID NO: 10, SEQ ID NO: 11 or SEQ ID NO: 15.
  • the polypeptide is a mutated HNL4X or a mutated HNL4Y according to the present invention, it has the SEQ ID NO: 9 or a sequence of at least 20, at least 50 and at least 100 consecutive amino acids or more derived from SEQ ID NO: 9.
  • polypeptide is defined as being any peptide or protein comprising at least two amino acids linked by a modified or unmodified peptide link.
  • polypeptide refers to molecules of short chains such as peptides, oligopeptides or oligomers or to long chains such as proteins.
  • a polypeptide according to the present invention can comprise modified amino acids.
  • the polypeptide of the present invention can also be modified by a natural method such as post-transcriptional modifications or by a chemical method.
  • any modification of the polypeptide which does not have the effect of eliminating the biochemical characteristics of the original polypeptide, i.e. the ability to form functional synapses, is covered within the scope of the present invention.
  • the invention relates to an isolated or purified polynucleotide coding for a polypeptide as defined above and more particularly to an isolated or purified polynucleotide coding for a polypeptide involved in synaptogenesis in which at least one mutation of this polynucleotide is associated with development of neurological diseases and / or a predisposition to the development of mental illnesses or psychiatric illnesses.
  • isolated or purified is meant the molecules which have been altered by humans from their native state, that is to say, if such a molecule exists in nature, it has been changed and / or removed from its initial environment.
  • a polynucleotide or polypeptide naturally present in a living organism is not “isolated”.
  • the same polynucleotide or polypeptide when separated from its normal environment and / or obtained by cloning, amplification and / or by chemical synthesis is considered according to the present invention as being “isolated”.
  • a polynucleotide or polynucleotide which is introduced into an organism by transformation, genetic manipulation or by any other method of recombination is “isolated” even if it is present in said organism.
  • polynucleotide is meant any DNA or RNA sequence or molecule having two or more nucleotides, including the nucleic sequences encoding an entire gene.
  • polynucleotide encompasses all nucleic acid molecules that are found in a natural or artificial state. This includes DNA molecules, RNA molecules, cDNAs, expressed sequences (ESTs), artificial sequences and all fragments thereof. It goes without saying that the definitions "derivative”, “variant” and “mutated” also apply to the polynucleotides according to the present invention. Any polynucleotide which has been chemically, enzymatically or metabolically modified but which has retained the biochemical properties of the original polypeptide, that is to say which has retained its power to form functional synapses, is included within the scope of the present invention. .
  • the polynucleotide according to the invention when it codes for an HNL3 protein or a fragment of this protein, the latter advantageously comprises SEQ ID NO: 14.
  • the polynucleotide comprises a sequence chosen from the group consisting of: SEQ ID NO: 12, and the sequences of at least 20, at least 50 and at least 100 consecutive or more nucleotides derived from SEQ ID NO: 12.
  • the polynucleotide according to the invention when it codes for an HNL4X or HNL4Y protein or a fragment of this protein, the latter advantageously comprises SEQ ID NO: 3, SEQ ID NO: 6, SEQ ID NO: 8.
  • the polynucleotide comprises a sequence chosen from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 16, SEQ ID NO: 17, and the sequences of at least 20, at least 50 and at least 100 consecutive nucleotides or more derived from these sequences.
  • the polynucleotide codes for a non-functional mutated protein.
  • the polynucleotide codes for a mutated HNL3 or mutated HNL4X protein.
  • the polynucleotide is mutated so that the mutation causes early termination of the protein.
  • the polynucleotide of the invention comprises SEQ ID NO: 1 and the mutation is an insertion of a thymine at position 1186 from position 465 of Figure 9 (ORF). This mutation causes the production of a defective protein devoid of its transmembrane part since this mutation causes early termination of the protein (D396stop).
  • the mutation causes a modification of the sequence protein such as an amino acid substitution at position 451 and / or 796 in Figure 18 or 21. More specifically, the mutation produced at position 451 consists in the substitution of an arginine by a cysteine, while the mutation produced at position 796 consists of the substitution of an asparagine by a serine.
  • This amino acid, arginine R451 is located in the acetylcholine esterase domain of neuroligins and is extremely conserved in all neuroligins sequenced to date and in the acetylcholine esterases of fish, birds and reptiles ( Figures 6A and 6B).
  • polypeptides and polynucleotides according to the present invention can be prepared by any suitable method. They can in particular be obtained by chemical synthesis, but it is also possible to obtain them by biological means, in particular by using different vectors in appropriate cell cultures as will be described below.
  • the peptides according to the present invention can be in deglycosylated, or glycosylated form, if necessary. A person skilled in the field of the invention will be able to obtain different polynucleotides / polypeptides and he will also be able to determine which of the polynucleotides / polypeptides obtained have those which have adequate biological activity.
  • the invention relates to any vector (cloning and / or expression) and any cellular host (prokaryotic or eukaryotic) transformed by such a vector, and comprising the regulatory elements allowing expression of the nucleotide sequence coding for a peptide according to the invention.
  • the subject of the invention is a process for the preparation of a peptide of the invention, by transformation of a cellular host using an expression vector (plasmid, cosmid, virus, etc. .) comprising the DNA sequences coding for the peptides of the invention, followed by culturing the cell host thus transformed, and recovering the peptide from the culture medium.
  • an expression vector plasmid, cosmid, virus, etc. .
  • the use of vectors for the expression of proteins and peptides in the cells of a host, in particular the human, is well known and will not be described in more detail.
  • polypeptides and polynucleotides of the present invention can also be used to prepare polyclonal or monoclonal antibodies capable of binding (preferably specifically) to at least one polypeptide / polynucleotide object of the invention.
  • the present invention therefore also relates to such purified antibodies which can be obtained by very well known techniques such as example the technique described by Kolher and Milstein (Continuous cultures of fused cells secreting antibody of predefined specificity, Nature, 1975, 262: 495-497).
  • the antibodies are of the “humanized” type. A person skilled in the art through his general knowledge will know how to prepare these types of antibodies.
  • vector refers to a polynucleotide construct designed to be transfected into different cell types.
  • these vectors target the expression vectors designed for the expression of a nucleotide sequence in a host cell; cloning vectors designed for the isolation, propagation and replication of inserted nucleotides; viral vectors designed for the production of recombinant virus or viral-like particle; or shuttle vectors that include attributes from more than one vector.
  • the invention relates to the treatment or prevention of biochemical pathologies or mental illnesses such as autism or Asperger's Syndrome. More particularly, the invention relates to the use of a non-mutated polynucleotide encoding a protein involved in synaptogenesis.
  • the protein consists of a cell adhesion protein, more preferably a protein belonging to the family of human neuroligins and even more preferably the polypeptide consists of the protein HNL3, HNL4X or the protein HNL4Y. Examples of non-mutated polynucleotides are given above.
  • the invention also relates to a treatment method comprising the insertion into at least a portion of the cells of a sick patient, of a polynucleotide encoding a polypeptide involved in synaptogenesis such as the protein HNL3 or HNL4X.
  • a polynucleotide encoding a polypeptide involved in synaptogenesis such as the protein HNL3 or HNL4X.
  • the cells into which the polynucleotide is inserted are stem cells. Examples of satisfactory polynucleotides are given above.
  • the invention relates to a method of transforming stem cells of a patient having a mutation of a gene coding for a protein involved in synaptogenesis, the method comprising: a) the use of stem cells of the patient ; b) insertion into the stem cell genome of a polynucleotide encoding a functional polypeptide involved in synaptogenesis such as the protein HNL3 or HNL4X; and c) reimplantation in the patient of cells transformed according to step b).
  • a person skilled in the art will be able to adapt the above-mentioned methods of treatment and determine, depending on several factors, the polynucleotides to be used, the means of inserting them into cells and the mode and quantity of polynucleotides or cells. to be administered.
  • factors that can influence his choices are: the nature of the treatment, the exact sequence of the polynucleotides; the stage of the disease; the patient's condition, age and weight, etc.
  • the invention also relates to a method for detecting biochemical disorders altering the formation, stabilization and / or recognition of synapses, a predisposition to the development of psychiatric pathologies and / or a mental illness such as autism. or Asperger's Syndrome.
  • the method comprises at least one of the following steps: the detection of a mutation in the sequence of a gene coding for a protein involved in synaptogenesis, in the sequence of a fragment of this gene or in the sequence of a messenger RNA of this gene; - detecting the presence of a protein involved in synaptogenesis; detecting a mutation in a protein involved in synaptogenesis; measuring the biological activity of a protein involved in synaptogenesis or its interaction with one of its protein partners.
  • a method for measuring such an interaction is for example described in Ichtchenko et al. (J. Biol. Chem., 1996, 271 (5): 2676-82) or Grifman et al. (Proc. Natl. Acad. Sci.
  • the method comprises: a) amplification of a gene coding for a protein involved in synaptogenesis, amplification of a fragment of said gene or amplification of a messenger RNA of said gene; and b) detecting a mutation in the sequence of said gene, in the sequence of said fragment or in the sequence of said messenger RNA.
  • kit for the detection of biochemical disorders altering the formation of synapses, of a predisposition to the development of psychiatric pathologies and / or of a mental illness, and / or for the diagnosis of mental illness.
  • the kit comprises at least one of the elements chosen from the group consisting of: a probe, an antibody, a reagent and a solid support, these elements allowing: i) the detection of a mutation in the sequence a gene coding for a protein involved in synaptogenesis, in the sequence of a fragment of this gene or in the sequence of a messenger RNA of this gene; and / or ii) measuring the biological activity of a protein involved in synaptogenesis or its interaction with one of its protein partners.
  • the elements chosen from the group consisting of: a probe, an antibody, a reagent and a solid support, these elements allowing: i) the detection of a mutation in the sequence a gene coding for a protein involved in synaptogenesis, in the sequence of a fragment of this gene or in the sequence of a messenger RNA of this gene; and / or ii) measuring the biological activity of a protein involved in synaptogenesis or its interaction with one of its protein partners.
  • the gene to which reference is made in the method and the kit codes, in its wild form, for a cell adhesion protein, more preferably for a protein belonging to the family of human neuroligins and even more preferably for the protein HNL3. , HNL4X or the HNL4Y protein.
  • the gene codes, in its wild form, for a protein comprising SEQ ID NO: 3, SEQ ID NO: 6, SEQ ID NO: 8 or SEQ ID NO: 14. More preferably, the gene comprises SEQ ID NO: 1, SEQ ID NO: 4 or SEQ ID NO: 12.
  • the invention relates to a method for sorting molecules which can make it possible to modulate the biological activity of a polypeptide encoded by the polynucleotide as defined above, or the biological activity of the polypeptide as defined above.
  • the sorting process comprises: a) bringing said polypeptide into contact with a molecule capable of modulating its biological activity; b) measuring the biological activity of said polypeptide; and c) evaluating the activity measured in step b) relative to a measurement of the biological activity of said polypeptide in the absence of said molecule.
  • the present invention also relates to the use of these polypeptides and polynucleotides encoding them for the preparation of therapeutic compositions. useful in the treatment of a mental or neurological disease, such as autism, Asperger's syndrome, schizophrenia or ADHD syndrome.
  • the composition of the present invention further contains a pharmaceutically acceptable vehicle, and an element selected from the group consisting of: a polynucleotide according to the present invention; - a polypeptide according to the present invention; an antibody according to the present invention; a vector according to the present invention; and - a host cell according to the present invention.
  • compositions according to the present invention can be in any solid or liquid form customary for pharmaceutical administration, that is to say for example forms of liquid administration, in gel, or any other support allowing for example the controlled release.
  • compositions which can be used mention may in particular be made of injectable compositions more particularly intended for injections into the blood circulation in humans.
  • a person skilled in the art will know how to prepare pharmaceutically acceptable compositions and to determine, according to several factors, the preferred mode of administration and the quantity to be administered. Among the factors that can influence his choices are: the nature of the treatment, the exact nature of the ingredients, active or not, used in the composition; the stage of the disease; the patient's condition, age and weight, etc.
  • HNL4X human neuroligin 4 gene
  • HNL1 HNL1
  • HNL2 chromosomes 3q26
  • HNL3 HNL3
  • HNL4X Xp22.3
  • HNL4Y Yq11.2
  • This table groups together the synonymous (KS) and non-synonymous (KA) 5 substitution rates of all known genes on the X chromosome having a homolog on the Y chromosome.
  • the KS / KA ratio is an indication of gene conservation.
  • KS is the synonymous substitution rate per synonymous site which represents the modifications which do not change the sequence of the protein.
  • the isolation of the HNL4X and HNL4Y genes was carried out by computer analysis of the sequences of the Xp22.3 and Yq11.22 region and by amplification of the complete transcripts from brain mRNAs.
  • Computer analysis A systematic study of the genes of the Xp22.3 region, close to the DXS996 microsatellite, was carried out using data from the sequencing of the human genome (http://genome.usc.edu and http // www. Ensembl.org / genomecentral).
  • the DXS996 microsatellite is the genetic marker that shows the most significant link with autism in the analysis by Philippe et al. (1999, supra).
  • the complete cDNAs of the HNL4X and HNL4Y mRNAs were back-transcribed, amplified and directly sequenced.
  • the oligonucleotides used for amplification and sequencing are indicated in Tables 2 and 3.
  • Table 3 Names and sequences of the primers used to sequence the HNL4X and HNL4Y genes
  • HNLXYE1R CACGGGAAAGGGGTGCATGGA 29
  • HNLXYE1R CACGGGAAAGGGGTGCATGGA 31
  • Table 4 Names and sequences of the primers used for the amplification of the MNL4 cDNA (mouse 57BL6)
  • Table 5 Names and sequences of the primers used for the amplification of MNL4 in three PCRs of approximately 1 kb
  • HNL4X / 4Y influences the synaptogenesis and mutation of HNL4X / 4Y constitutes a factor. predisposition to mental illness, including autism and Asperger's syndrome.
  • This Stop mutation in a specific gene for primates, carried by the X chromosome in two autistic subjects and involved in synaptogenesis is one of the first functional mutations identified in a psychiatric illness. This mutation is also the first mutation described associated with autism without any other clinical sign (fragile X, tuberculous sclerosis, etc.).
  • Example 2 Characterization of the HNL3 gene and its implication in psychiatric syndromes.

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EP02801090A 2001-11-30 2002-12-02 Polynukleotid, protein und varianten, verwickelt in die synaptogenese, und deren diagnostische und therapeutische anwendungen Withdrawn EP1453861A2 (de)

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CA2364106 2001-11-30
CA002364106A CA2364106A1 (fr) 2001-11-30 2001-11-30 Polynucleotide et proteine impliques dans la synaptogenese, variants de ceux-ci, et leurs applications therapeutiques et diagnostiques
PCT/FR2002/004134 WO2003045998A2 (fr) 2001-11-30 2002-12-02 Polynucléotide et protéine impliqués dans la synaptogenèse, variants de ceux-ci, et leurs applications thérapeutiques et diagnostiques.

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DE602004021186D1 (de) * 2003-08-22 2009-07-02 Integragen Sa Menschliches autismus-suszeptibilitätsgen und verwendungen davon
CA2523399A1 (en) * 2005-10-14 2007-04-14 Institut Pasteur Genetic variations associated with psychiatric disorders
US8008259B2 (en) 2005-11-07 2011-08-30 Copenhagen University, Techtrans Unit Neurotrophin-derived peptide sequences
AU2016204711B2 (en) * 2008-10-01 2018-03-01 Immatics Biotechnologies Gmbh Novel immunotherapy against several tumors including neuronal and brain tumors
RS53782B1 (sr) 2008-10-01 2015-06-30 Immatics Biotechnologies Gmbh Preparati tumor-asociranih peptida i odgovarajuća antikancerska vakcina za tretman glioblastoma (gbm) i drugih kancera
AU2014271235B2 (en) * 2008-10-01 2017-03-02 Immatics Biotechnologies Gmbh Novel immunotherapy against several tumors including neuronal and brain tumors
WO2011112961A1 (en) * 2010-03-12 2011-09-15 Children's Medical Center Corporation Methods and compositions for characterizing autism spectrum disorder based on gene expression patterns
US20130058871A1 (en) * 2011-07-28 2013-03-07 Howard Hughes Medical Institute Method and system for mapping synaptic connectivity using light microscopy
WO2018106982A1 (en) * 2016-12-09 2018-06-14 The Regents Of The University Of California Compositions and methods for enhancing beta cell maturation, health and function

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AU3773099A (en) * 1998-04-29 1999-11-16 Government Of The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services, The Identification of polymorphisms in the pctg4 region of xq13

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US20090117554A1 (en) 2009-05-07
AU2002364808A1 (en) 2003-06-10
WO2003045998A3 (fr) 2004-04-22
AU2002364808A8 (en) 2003-06-10
US7384740B2 (en) 2008-06-10
CA2364106A1 (fr) 2003-05-30
WO2003045998A2 (fr) 2003-06-05
US20050118588A1 (en) 2005-06-02
US7906640B2 (en) 2011-03-15
US20090202992A1 (en) 2009-08-13

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